Isolating DNA from tissues and paraffin-embedded tissue samples can be challenging as double-stranded DNA is physically fragile and highly susceptible to exo- and endonucleases. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in the presence of DNAse inhibitors. Further, extracting DNA from the nucleus need specific methods by combining physical, mechanical and chemical lysis approaches,
Get tips on using jetPEI® DNA transfection, HTS application to perform DNA transfection Mammalian cells - Primary cells Rat aortic smooth muscle cells (rASMC)
Stem cells have the unique ability to self-renew or differentiate themselves into various cell types in response to appropriate signals. These cells are especially important for tissue repair, regeneration, replacement, or in the case of hematopoietic stem cells (HSCs) to differentiate into various myeloid populations. Appropriate signals refer to the growth factor supplements or cytokines that mediate differentiation of various stem cells into the required differentiated form. For instance, HSCs can be differentiated into dendritic cells (with IL-4 and GM-CSF), macrophages (with m-CSF) and MDSCs (with IL-6 and GM-CSF). Human pluripotent stem cells (hPSCs) and induced pluripotent stem cells (iPSCs) can be first cultured in neural differentiation media (GSK3𝛃-i, TGF𝛃-i, AMPK-i, hLIF) to form neural rosettes, which can be differentiated into neural or glial progenitors (finally differentiated into oligodendrocytes). Neural progenitors can be finally differentiated into glutaminergic (dibytyryl cAMP, ascorbic acid) and dopaminergic (SHH, FGF-8, BDNF, GDNF, TGF-𝛃3) neurons. Thus, it is important to first identify the self-renewing cell line: its source and its final differentiation state, followed by the supplements and cytokines required for the differentiation, and final use. Timelines are another thing that is considered. For instance, it takes 7-10 days to form neural rosettes from iPSCs and 3 days to differentiate neural progenitors to neurons. Finally, the stability for stem cell culture media varies. It is advised to make fresh media every time when differentiating HSCs to myeloid populations, whereas neural differentiation media may remain stable for two weeks when stored in dark between 2-8C.
Get tips on using Silencer® FANCD2 siRNA (human) to perform siRNA / miRNA gene silencing Human - 501 Mel and SK Mel 28 FANCD2
Get tips on using FuGENE® HD Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat microglia
Get tips on using Lipofectamine® 2000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat microglia
Get tips on using Lipofectamine™ 3000 Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Rat astrocytes
Get tips on using siGENOME Human ARAP1 (116985) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Human - HeLa ARAP1
Get tips on using siGENOME Human FTO (79068) siRNA - Individual to perform siRNA / miRNA gene silencing Human - SHSY5Y FTO
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